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目的 :建立大肠癌细胞中提取和测定 5-Fu含量的 HPL C方法 ,并观察药物浓度、时间及给药次数与大肠癌细胞内 5-Fu含量的关系。方法 :用 0 .1%氢氧化钠裂解大肠癌细胞后用乙醚-异丙醇 ( 8∶ 2 )提取细胞内 5-Fu,HPL C法测定大肠癌细胞内 5-Fu含量。分析条件 :Spherisorb C18柱 ,流动相 :0 .0 5m ol/L 磷酸盐缓冲液 ,紫外检测波长 :2 65nm。结果 :5-Fu进样量在 0 .0 0 0 52 6μg～ 0 .42 0 8μg之间呈良好线性关系 ,线性方程为 Y=3 154159X-40 8.89,r=0 .9997;5-Fu的最小检测限为 5.2 6ng;5-Fu不同加入量的的平均回收率分别为 82 .15%和 87.50 % ;重复性试验 RSD=6.72 %。结论 :用 HPL C法测定大肠癌细胞中 5-Fu含量的方法是可行的 ;大肠癌细胞对 5-Fu的摄取可能存在“饱和现象”,当给药浓度达到一定量 ( 5μg/m L)时 ,癌细胞中 5-Fu含量最高。
OBJECTIVE: To establish a method for extracting and assaying 5-Fu content in colorectal cancer cells, and to observe the relationship between concentration and time of drug and the number of 5-Fu in colorectal cancer cells. METHODS: Colorectal cancer cells were lysed with 0.1% sodium hydroxide and intracellular 5-Fu was extracted with ether-isopropanol (8: 2). The 5-Fu content was determined by HPL-C method. Analysis conditions: Spherisorb C18 column, mobile phase: 0. 05mol / L phosphate buffer, UV detection wavelength: 2 65nm. Results: The 5-Fu injection volume showed a good linear relationship between 0 0 0 526μg ~ 0. 42 0 8μg, the linear equation was Y = 3 154159X-40 8.89, r = 0.9997; The minimum detectable limit was 5.26ng. The average recoveries of 5-Fu at different loadings were 82.15% and 87.50%, respectively. The repeatability test RSD was 6.72%. Conclusion: It is feasible to determine the 5-Fu content in colorectal cancer cells by HPL-C method. The uptake of 5-Fu in colorectal cancer cells may be “saturated”. When the concentration of 5-Fu is up to 5μg / When the cancer cells in the highest 5-Fu content.